Cascino J J, Harris R F, Smith C S, Andrews J H
Departments of Botany, Soil Science, and Plant Pathology, University of Wisconsin-Madison, Madison, Wisconsin 53706.
Appl Environ Microbiol. 1990 Aug;56(8):2303-2310. doi: 10.1128/aem.56.8.2303-2310.1990.
The effect of V8 juice concentration (5 to 40%, vol/vol), spore inoculum density (10 and 10 spores per ml), and liquid batch or fed-batch culture condition on mycelium and spore production by Colletotrichum gloeosporioides was evaluated. The amount of mycelium produced, the time required for initiation of sporulation following attainment of maximum mycelium, and the time for attainment of maximum spore concentration increased with increasing V8 juice concentration in batch culture. Cultures containing V8 juice at >10% achieved a similar spore density (apparent spore-carrying capacity) of about 0.8 mg of spores per ml (1 x 10 to 2 x 10 spores per ml) independent of inoculum density and V8 juice concentration. The relative spore yield decreased from a high of 64% of the total biomass for the low-inoculum 5% V8 culture, through 13% for the analogous 40% V8 culture, to a low of 2% for the high-inoculum 27% V8 culture. Fed-batch cultures were used to establish conditions of high spore density and low substrate availability but high substrate flux. The rate of addition of V8 juice was adjusted to approximate the rate of substrate utilization by the (increasing) biomass. The final spore concentration was about four times higher (3.0 mg of spores per ml) than the apparent spore-carrying capacity in batch culture. This high spore yield was obtained at the expense of greatly reduced mycelium, resulting in a high relative spore yield (62% of the total biomass). Microcycle conidiation occurred in the fed-batch but not batch systems. These data indicate that substrate-limited, fed-batch culture can be used to increase the amount and efficiency of spore production by C. gloeosporioides by maintaining microcycle conidiation conditions favoring allocation of nutrients to spore rather than mycelium production.
评估了V8果汁浓度(5%至40%,体积/体积)、孢子接种密度(每毫升10和10个孢子)以及液体分批或补料分批培养条件对胶孢炭疽菌菌丝体和孢子产生的影响。在分批培养中,随着V8果汁浓度的增加,产生的菌丝体量、达到最大菌丝体后开始产孢所需的时间以及达到最大孢子浓度的时间均增加。含有大于10% V8果汁的培养物,无论接种密度和V8果汁浓度如何,都能达到相似的孢子密度(表观孢子承载能力),约为每毫升0.8毫克孢子(每毫升1×10至2×10个孢子)。相对孢子产量从低接种量5% V8培养物中占总生物量的64%的高位,下降到类似的40% V8培养物中的13%,再到高接种量27% V8培养物中的2%的低位。补料分批培养用于建立高孢子密度、低底物可用性但高底物通量的条件。V8果汁的添加速率被调整为接近(不断增加的)生物量对底物的利用速率。最终孢子浓度比分批培养中的表观孢子承载能力高约四倍(每毫升3.0毫克孢子)。这种高孢子产量是以菌丝体大幅减少为代价获得的,从而导致相对孢子产量较高(占总生物量的62%)。微循环产孢发生在补料分批培养系统中,而不是分批培养系统中。这些数据表明,底物受限的补料分批培养可用于通过维持有利于将营养物质分配到孢子而非菌丝体生产的微循环产孢条件,来增加胶孢炭疽菌孢子产生的量和效率。